NR4A1过表达对APP代谢和Tau磷酸化影响的研究

发布时间：2018-05-17 00:20

本文选题：NR4A1 + APP　；参考：《重庆医科大学》2017年硕士论文

【摘要】：研究目的:探讨孤儿核受体NR4A1过表达对APP代谢和Tau磷酸化的影响,并阐述其可能机制。研究方法:18月龄APP/PS1和WT小鼠麻醉后处死取海马组织。应用蛋白质印迹法检测NR4A1蛋白质的表达,实时定量聚合酶链反应(q RT-PCR)检测NR4A1 mRNA表达,免疫组化染色观察NR4A1在小鼠大脑海马中的表达。用鼠pCMV-NR4A1质粒转染HT22细胞,应用蛋白质印迹法检测NR4A1、APP、ADAM10、BACE1、t-Tau和p-Tau、GSK3β和p-GSK3β、CDK5、ERK的表达,q RT-PCR检测NR4A1 mRNA、ADAM10 mRNA、BACE1 mRNA表达。研究结果:APP/PS1小鼠海马组织中APP、NR4A1表达增多,q RT-PCR显示NR4A1 mRNA水平也升高。用鼠pCMV-NR4A1质粒转染HT22细胞后,NR4A1蛋白质及mRNA水平均显著升高。NR4A1过表达后,APP、BACE1表达升高,ADAM10表达下降;q RT-PCR显示NR4A1过表达后ADAM10 mRNA水平下降,BACE1 mRNA水平升高。NR4A1过表达后p-Tau(S396)蛋白表达升高,但是总的Tau及p-Tau(S262和T231)却没有改变;p-GSK3β(S9)表达下降,CDK5、ERK表达无明显改变。研究结论:综上可知,NR4A1过表达使ADAM10下调、BACE1上调,从而促进APP向Aβ途径降解;此外,NR4A1可能通过p-GSK3β影响Aβ和p-Tau的生成。
[Abstract]:Aim: to investigate the effects of NR4A1 overexpression on APP metabolism and Tau phosphorylation in orphan nuclei and to elucidate its possible mechanism. Methods the hippocampal tissues of APP/PS1 and WT mice were killed after anesthesia at 18 months old. The expression of NR4A1 protein was detected by Western blot, the expression of NR4A1 mRNA was detected by real-time quantitative polymerase chain reaction (PCR), and the expression of NR4A1 in the hippocampus of mice was observed by immunohistochemical staining. HT22 cells were transfected with mouse pCMV-NR4A1 plasmid. The expression of NR4A1 mRNA-ADAM10 mRNA-BACE1 mRNA was detected by Western blotting. The expression of NR4A1 mRNA-ADAM10 mRNA-BACE1 mRNA and p-Tautaug GSK3 尾 and p-GSK3 尾 CDK5 ERK were detected by Western blot. Results the increased expression of APPtNR4A1 in hippocampal tissue of mice with 1: App / PS1 showed that the level of NR4A1 mRNA was also increased by Q RT-PCR. After transfection of mouse pCMV-NR4A1 plasmid into HT22 cells, the levels of NR4A1 protein and mRNA were significantly increased. After overexpression of NR4A1, the expression of APPfBACE1 increased and the expression of ADAM10 decreased. Q RT-PCR showed that the level of ADAM10 mRNA decreased after NR4A1 overexpression, and the expression of p-TauS396) protein increased after overexpression of NR4A1, and the expression of p-TauS396 protein increased after overexpression of NR4A1. However, total Tau, p-Tau(S262 and T231) did not change the expression of p-GSK3 尾 -S9). Conclusion: the overexpression of NR4A1 leads to down-regulation of ADAM10 and up-regulation of APP to A 尾 pathway, and NR4A1 may affect the formation of A 尾 and p-Tau through p-GSK3 尾.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749.16

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